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3T3-L1 Cells

3T3-L1 cells are a well-established mouse embryonic fibroblast cell line that can differentiate into adipocytes (fat cells) under appropriate culture conditions.
These cells have been widely used as a model system to study adipocyte biology, including adipogenesis, lipid metabolism, and insulin signaling. 3T3-L1 cells are known for their ability to rapidly proliferate and undergo differentiation in response to hormonal stimuli, making them a valuable tool for researchers investigating the cellular and molecular mechanisms underlying adipose tissue development and function.
The 3T3-L1 cell line is an importatnt model for the study of obesity, diabetes, and other metabolic disorders.

Most cited protocols related to «3T3-L1 Cells»

1
Adipocyte Differentiation by BMPs
To induce adipocyte differentiation by BMPs in the absence of induction cocktails, both WT brown preadipocytes and 3T3-L1 white preadipocytes were grown in regular growth medium supplemented with combination of rhBMPs (3.3 to 8.3 nM), insulin (20 nM) and T3 (1 nM) or vehicle as indicated in the text and figure legends for 7–13 days. To stimulate thermogenic program, differentiated cells were incubated with 500 µM dibutyrul cyclic AMP for 4 hrs. Cells were grown in growth medium without hormonal supplements for 18 hrs prior to cAMP stimulation.
C3H10T1/2 cells were grown in the presence and absence of 8.3 nM rhBMP-7 for 3 days to reach confluence (day 3). These cells were then induced to adipocyte differentiation using protocols described below for additional 7 days (day 10). Adipocyte differentiation was done by treating confluent cells for 48 hours in medium supplemented with 20 nM insulin and 1 nM triiodothyronine (T3), 0.5 mM isobutylmethylxanthine (IBMX), 5 µM dexamethasone, and 0.125 mM indomethacin. Cells were placed back to growth medium supplemented with insulin and T3, which was then changed every second day. After four to five more days in this medium, cells exhibited a fully differentiated phenotype with massive lipid accumulation.
Tseng Y.H., Kokkotou E., Schulz T.J., Huang T.L., Winnay J.N., Taniguchi C.M., Tran T.T., Suzuki R., Espinoza D.O., Yamamoto Y., Ahrens M.J., Dudley A.T., Norris A.W., Kulkarni R.N, & Kahn C.R. (2008). New role of bone morphogenetic protein 7 in brown adipogenesis and energy expenditure. Nature, 454(7207), 1000-1004.
Publication 2008
3T3-L1 Cells Adipocytes Bone Morphogenetic Proteins Cells Cyclic AMP Dexamethasone Indomethacin Insulin Liothyronine Lipid A Phenotype Thermogenesis
2
Proteome Analysis of Kaempferol Treatment
Cell lysates were obtained from 3T3-L1 or HepG2 cells. Cells were scraped and lysed with M-PER lysis buffer. After centrifugation for 15 min at 16,000×g, the supernatant was obtained, and protein content was quantified using Bradford reagent. Before drug treatment, the samples were diluted to achieve a protein concentration of 1 mg/mL. Samples were treated with the Kaem or DMSO for 2 h at 25 °C and then incubated with pronase (5, 10, and 20 µg/mL) or distilled water for 10 min at 25 °C. After the reaction, SDS was added to the sample and the samples were heated at 100 °C. A portion of each sample was used for LC–MS/MS analysis. Sample preparation and proteome analysis were conducted as indicated in the previous publication69 . For western blot analysis, VDAC1 or Na+K+ ATPase was used as an internal control. For the structure–activity-relationship (SAR) analysis, kaempferol (Sigma-Aldrich, 60010), Acacetin (Sigma-Aldrich, 00017), isosakuranetin (Sigma-Aldrich, PHL82569), Biochanin A (Sigma-Aldrich, D2016), (−)Epicatechin (Sigma-Aldrich, E4018), Genistein (Sigma-Aldrich, G6649) were used.
Kim D., Hwang H.Y., Ji E.S., Kim J.Y., Yoo J.S, & Kwon H.J. (2021). Activation of mitochondrial TUFM ameliorates metabolic dysregulation through coordinating autophagy induction. Communications Biology, 4, 1.
Publication 2021
3T3-L1 Cells acacetin ATP8A2 protein, human biochanin A Buffers Cells Centrifugation Epicatechin Genistein Hep G2 Cells isosakuranetin kaempferol Na(+)-K(+)-Exchanging ATPase Pharmaceutical Preparations Pronase Proteins Proteome Sulfoxide, Dimethyl Tandem Mass Spectrometry VDAC1 protein, human Western Blot
3
Fluorescence Imaging of Lysosomal and Autophagic Markers
For acridine orange (AO) stain in the screen, HeLa cells were grown in 96-well plates. Cells were treated with DMSO or indatraline, bafilomycin A1, and 658-natural chemicals for 24 h, and stained with 5 μg/mL AO. Fluorescence intensity was measured by victor plate reader, where fluorescence intensity of each well in the plate is promptly displayed as numerical readout. For confocal microscopy, 3T3-L1 or HeLa cells were grown on 15 mm coverslips at a density of 1.0 × 105 cells/well in 6-well plates The cells were then treated with drugs for the time periods indicated, followed by treatment with 5 μg/mL AO (Sigma-Aldrich). Nuclei were stained with Hoechst. Following incubation for 20 min, the cells were fixed with 4% PFA and washed three times with PBS. Images were obtained using an LSM880 confocal microscope at ×400 magnification. Red fluorescence intensity was quantified using Image J2 software. For BODIPY FL-Pepstatin A stain, HeLa cells were grown on 15 mm coverslips at a density of 1.0 × 105 cells/well in 6-well plates. The cells were then treated with drugs for the time periods indicated, followed by treatment with 1 μM BODIPY FL-Pepstatin A (Invitrogen) for 30 min. Nuclei were stained with Hoechst. Following incubation for 20 min, the cells were fixed with 4% PFA and washed three times with PBS. Images were obtained using an LSM880 confocal microscope at 400× magnification. For DQ-BSA analysis, HeLa cells were grown on 15 mm coverslips at a density of 1.0 × 105 cells/well in 6-well plates. The cells were then treated with 10 μg/mL DQ-BSA for 2 h. After change medium, cells were treated with drugs for the time periods indicated. Nuclei were stained with Hoechst with incubation for 20 min, the cells were fixed with 4% PFA and washed three times with PBS. Images were obtained using an LSM880 confocal microscope at ×400 magnification.
Kim D., Hwang H.Y., Ji E.S., Kim J.Y., Yoo J.S, & Kwon H.J. (2021). Activation of mitochondrial TUFM ameliorates metabolic dysregulation through coordinating autophagy induction. Communications Biology, 4, 1.
Publication 2021
3T3-L1 Cells Acridine Orange bafilomycin A1 BODIPY Cell Nucleus Cells Fluorescence HeLa Cells indatraline Microscopy, Confocal pepstatin Pharmaceutical Preparations Sulfoxide, Dimethyl
4
Adipocyte Differentiation Assays Protocols
For adipocyte differentiation assays, confluent cultures of 3T3-L1 and Swiss 3T3 subclones were exposed to induction medium containing dexamethasone (1 μM), insulin (5 μg/ml), and isobutylmethylxanthine (0.5 mM) (DMI) and 10% FBS. 48 hours after induction, cells were maintained in DMEM containing insulin (5 μg/ml) and 10% FBS until ready for harvest. For NIH 3T3 cells, differentiation medium contained DMI, 6% FBS, and 1 μM rosiglitazone. After induction, cells were maintained in medium containing 6% FBS, insulin, and 1 μM rosiglitazone until ready for harvest. For BMP-induced adipogenesis of NIH 3T3 cell lines, cells were grown to confluence, and maintained at post-confluence, in medium containing 6 ng/mL BMP4 or 25 ng/mL BMP2 along with insulin and rosiglitazone. The derivation and genotyping of Zfp423 knockout mice has been previously described 14 (link), 23 (link). All animal experiments were performed according to procedures approved by the Dana-Farber Cancer Institute’s and Beth Isreal Deconess Medical Center’s Institutional Animal Care and Use Committee.
Gupta R.K., Arany Z., Seale P., Mepani R.J., Ye L., Conroe H.M., Roby Y.A., Kulaga H., Reed R.R, & Spiegelman B.M. (2010). Transcriptional Control of Preadipocyte Determination by Zfp423. Nature, 464(7288), 619-623.
Publication 2010
3T3-L1 Cells Adipocytes Adipogenesis Biological Assay BMP2 protein, human Bone Morphogenetic Protein 4 Cells Dexamethasone Institutional Animal Care and Use Committees Insulin Malignant Neoplasms Mice, Knockout NIH 3T3 Cells Rosiglitazone
5
Adipocyte Differentiation Assay
To induce differentiation, cells were seeded into 96-well black clear-bottom tissue culture plates (Greiner cat # 655090) at approximately 30,000 cells per well and grown to confluence in respective growth medium. Upon reaching confluency, cells were cultured for a further 48 hours in growth media to initiate growth arrest. After this window, to initiate clonal expansion and differentiation, media was replaced with test chemical/control dilution series (0.1 nM to 1.0 μM for RSG, TBT; 1 nM to 10 μM all other chemicals) in a 0.1% DMSO vehicle diluted in differentiation media (base media for OP9 and 3T3-L1 described above, supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, 1.0 μg/mL human insulin, and 0.5 mM 3-isobutyl-1-methylxanthine). For antagonist testing, cells were co-exposed to a test chemical dilution series (1 nM to 10 μM) as well as EC50 concentrations of rosiglitazone in each cell line: 9.0 nM, 30 nM, and 15 nM for ATCC 3T3-L1, Zenbio 3T3-L1, and OP9 cells, respectively. Following 48 hours of exposure, media was replaced with test chemicals and controls diluted in adipocyte maintenance media (differentiation media without 3-isobutyl-1-methylxanthine). This maintenance media (along with test chemicals and dilutions) was refreshed every 2–3 days until plates were assayed.
Kassotis C.D., Masse L., Kim S., Schlezinger J.J., Webster T.F, & Stapleton H.M. (2017). Characterization of Adipogenic Chemicals in Three Different Cell Culture Systems: Implications for Reproducibility Based on Cell Source and Handling. Scientific Reports, 7, 42104.
Publication 2017
3T3-L1 Cells Adipocytes Cell Lines Cells Clone Cells Culture Media Fetal Bovine Serum Homo sapiens Insulin methylxanthine Penicillins Rosiglitazone Streptomycin Sulfoxide, Dimethyl Technique, Dilution Tissues

Most recents protocols related to «3T3-L1 Cells»

1
Obesity and Adipocyte ACAT Expression
Not available on PMC !

Example 1

To examine the function of ACATs in obesity, the expression patterns of ACAT1 and ACAT2 genes, and their gene products during adipogenesis of murine 3T3-L1 preadipocytes in vitro were examined. ACAT1 mRNA level was markedly increased in adipocytes from 2 days after initiation of adipogenesis (i.e., D2) as judged by real-time PCR assay (FIG. 1). However, ACAT2 mRNA level was similar between preadipocytes (D0) and mature adipocytes (D6) while a temporal reduction of ACAT2 level was observed at D2 (FIG. 1). In addition, white adipose tissue (WAT) isolated from high fat diet-induced obese mice displayed elevated mRNA level of ACAT1 and reduced mRNA level of ACAT2 when compared with those in lean mice as judged by real-time PCR assay. Leptin level was measured in WAT from lean and obese mice to ensure the development of obesity (FIG. 2). In addition, brown adipose tissue (BAT) from obese mice exhibited elevated levels of both ACAT1 and ACAT2. Uncoupling protein-1 (UCP-1) level was measured in BAT from lean and obese mice as a BAT-specific marker protein (FIG. 2). However, liver from lean and obese mice exhibited similar levels of ACAT1 and ACAT2 (FIG. 2).

US11872198B2. Compositions and methods for regulating body weight and metabolic syndromes (2024-01-16). Purdue Research Foundation [US]. Inventors: Kee-Hong Kim [US], Yuyan Zhu [US].
Patent 2024
3T3-L1 Cells Adipocytes Adipogenesis a protein, mouse Biological Assay Brown Adipose Tissue Uncoupling Protein Brown Fat CES1 protein, human Diet, High-Fat Genes Leptin Liver Mice, Obese Mus Obesity Proteins Real-Time Polymerase Chain Reaction RNA, Messenger White Adipose Tissue
2
Culturing SiHa and 3T3-L1 Cells
Dulbecco's Modified Eagle Medium (DMEM) along with 10% heat-inactivated fetal bovine serum (FBS), 100 U mL−1 penicillin, and 100 μg mL−1 streptomycin were involved to culture SiHa cells (cancerous tissues of the cervix uteri) and 3T3-L1 (several normal cell lines of mouse). 310 K temperature was maintained during culture of the cells in a CO2 incubator with a 5% CO2 supply.
Acharya P., Kuila A., Pramanik U., Hathwar V.R., Brandao P., Mukherjee S., Maity S., Maity T., Maity R, & Chandra Samanta B. (2023). Combined theoretical and experimental insights on DNA and BSA binding interactions of Cu(ii) and Ni(ii) complexes along with the DPPH method of antioxidant assay and cytotoxicity studies. RSC Advances, 13(11), 7632-7644.
Publication 2023
3T3-L1 Cells Cell Culture Techniques Cell Lines Cervical Cancer Eagle Fetal Bovine Serum Mus Penicillins Streptomycin Tissues
3
Lipid Quantification in Adipocytes
Differentiated mature 3T3-L1 cells were given PA and PD for 24 h. The cells were gently rinsed with PBS before staining, followed by fixation with 4% paraformaldehyde for 1 h. Then cells were washed with PBS, and each well was stained with 2 mL of freshly configured Oil Red O working solution for 1–2 h. The staining solution was discarded, and 60% isopropanol was rinsed quickly once, followed by rinsing with ultrapure water three times, and observed under the microscope for photographs.
Lv Y., Lv X., Feng J., Cheng F., Yu Z., Guan F, & Chen L. (2023). (20R)-panaxadiol improves obesity by promoting white fat beigeing. Frontiers in Pharmacology, 14, 1071516.
Publication 2023
3T3-L1 Cells Cells Isopropyl Alcohol Microscopy paraform solvent red 27
4
3T3-L1 Preadipocyte Differentiation Protocol
3T3-L1 preadipocytes (ATTC, United States) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and 10% bovine calf serum (Gibco, United States) in an atmosphere of 5% CO2 at 37°C. For differentiation studies, 3T3-L1 cells were cultured in DMEM containing 10% FBS,1 μMOL dexamethasone (Sigma, United States), 0.5 mMOL IBMX (Sigma, United States) and 5 μg/mL insulin for 2 days and then replaced with DMEM culture medium without 10% FBS and 5 μg/mL insulin for 2 days. Then replaced with DMEM containing 10% FBS. DMEM cell culture medium containing 10% FBS was changed every 2 days until the 8–10th day.
Lv Y., Lv X., Feng J., Cheng F., Yu Z., Guan F, & Chen L. (2023). (20R)-panaxadiol improves obesity by promoting white fat beigeing. Frontiers in Pharmacology, 14, 1071516.
Publication 2023
1-Methyl-3-isobutylxanthine 3T3-L1 Cells Atmosphere Bos taurus Cells Culture Media Dexamethasone Eagle Insulin Serum
5
Anti-Adipogenic Activity Evaluation in 3T3-L1 Cells
The anti-adipogenic activity was assessed in the 3T3-L1 cell line using the methods explained by Pandeya et al22 (link) 3T3-L1 preadipocytes were cultured and subcultured in 10% NCS supplemented with DMEM growth medium with 1% antibiotics (penicillin–streptomycin). Cells were maintained under a humidified atmosphere of 5% CO2 at 37°C. Then, the obtained cells were seeded in a 96-well plate at a cell density of 7×103 cells per well and left overnight. The next day, the cells were treated with different concentrations of plant extracts in 10% FBS-supplemented DMEM. After 48 hours, the MTT assay was performed to calculate cell viability at different concentrations. Thus, a safe dose was used to evaluate the anti-adipogenic activity.
The Oil Red O (ORO) staining assay was used to quantify the amount of lipid deposited in differentiated 3T3-L1 cells. 3T3-L1 preadipocytes were seeded into the 24-well plate at a cell density of 5×103 cells per well in 10% FBS-supplemented DMEM culture medium. The cultured plates were kept in an incubator until cells reached full confluency, and the cell medium was changed every 2 days. After that, cells were co-treated with non-toxic concentrations of extracts (obtained by the MTT assay) in cell differentiation media for 2 days. The cell differentiation media contained dexamethasone (1 µM), insulin (5 µg/mL), and 3-isobutyl-2-methylxanthine (IBMX 0.5 mM) in 10% FBS-supplemented DMEM. Differentiation media with the tested concentrations of extracts were replaced after 2 days by adipocyte maintenance medium (10% FBS-supplemented DMEM with 5 µg/mL of insulin; without extracts). After 2 days, the adipocyte maintenance medium was replaced by 10% FBS-supplemented DMEM. On the eighth day of the experiment, cells were washed with phosphate-buffered saline and fixed in 10% formalin. Fixed cells were stained with ORO and viewed under an EVOS XL (Life Technologies, USA) microscope. The deposited stain was dissolved in isopropanol and quantified using a microplate reader at 520 nm.
The final results were expressed as percentage deposition of lipid in differentiated cells compared to control, as follows:
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\begin{document}
$$\rm Percentage\ lipid\ deposition=\left({Absorbance\ of\ sample/Absorbance\ of\ control} \right) \times100$$
\end{document}
Lamichhane G., Sharma G., Sapkota B., Adhikari M., Ghimire S., Poudel P, & Jung H.J. (2023). Screening of Antioxidant, Antibacterial, Anti-Adipogenic, and Anti-Inflammatory Activities of Five Selected Medicinal Plants of Nepal. Journal of Experimental Pharmacology, 15, 93-106.
Publication 2023
1-Methyl-3-isobutylxanthine 3-methylxanthine 3T3-L1 Cells Adipocytes Adipogenesis Antibiotics Atmosphere Biological Assay Cells Cell Survival Dexamethasone Formalin Insulin Isopropyl Alcohol Lipids Microscopy Penicillins Phosphates Plants Saline Solution solvent red 27 Stains Streptomycin

Top products related to «3T3-L1 Cells»

1
Dexamethasone (dex) by Merck Group
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Dexamethasone is a synthetic glucocorticoid medication used in a variety of medical applications. It is primarily used as an anti-inflammatory and immunosuppressant agent.
2
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
3
Insulin by Merck Group
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Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific
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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.
5
3t3 l1 preadipocyte by American Type Culture Collection
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3T3-L1 preadipocytes are a cell line derived from mouse embryonic fibroblasts. They are commonly used as a model for the study of adipocyte differentiation and metabolism.
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Penicillin streptomycin by Thermo Fisher Scientific
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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3t3 l1 cells by American Type Culture Collection
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3T3-L1 cells are an established mouse cell line derived from 3T3 cells. They are preadipocytes, capable of differentiating into mature adipocytes (fat cells) under the appropriate culture conditions.
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Oil red o by Merck Group
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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3-isobutyl-1-methylxanthine is a chemical compound primarily used as a research tool in laboratories. It functions as a nonselective phosphodiesterase inhibitor, which can affect various cellular processes. The core function of this product is to serve as a laboratory reagent for scientific research purposes.

More about "3T3-L1 Cells"

3T3-L1 cells are a widely-used mouse-derived embryonic fibroblast cell line that can differentiate into adipocytes (fat cells) under the right culture conditions.
These versatile cells have become a crucial model system for studying the cellular and molecular mechanisms underlying adipogenesis (fat cell formation), lipid metabolism, and insulin signaling pathways - key processes involved in conditions like obesity, diabetes, and other metabolic disorders.
The 3T3-L1 cell line is known for its rapid proliferation and robust differentiation capabilities when exposed to appropriate hormonal stimuli, making it an invaluable tool for researchers investigating adipose tissue development and function.
These preadipocytes can be induced to undergo adipocyte differentiation through the addition of a cocktail of adipogenic inducers, such as dexamethasone, insulin, and 3-isobutyl-1-methylxanthine (IBMX), in a defined culture medium like DMEM supplemented with fetal bovine serum (FBS) and penicillin/streptomycin.
During the differentiation process, the 3T3-L1 cells accumulate lipids, which can be visualized using lipid-specific stains like Oil Red O.
Researchers often utilize molecular biology techniques, such as TRIzol reagent for RNA extraction, to study the expression of adipogenic marker genes and signaling pathways that drive the transformation of fibroblasts into mature, functional adipocytes.
The 3T3-L1 cell model has been instrumental in advancing our understanding of adipose tissue biology and the underlying mechanisms of metabolic disorders, providing valuable insights for the development of potential therapeutic interventions.